The composition, biosynthesis and insertion of surface membrane glycoproteins and polypeptides in mammalian normal and cancer cells will be studied. Plasma membranes will be purified and the kinetics of synthesis of certain polypeptides and glycoproteins in the cells will be studied together with their insertion into the membrane. A membrane maturing cytopathic virus, vesicular stomatitis virus will be used to aid theses studies. Since a radioactive precursor (fucose) to glycoproteins will be used, the specific radioactivity and pool size of the soluble precursor pool (GDP-fucose) will be determined. This will enable us to compare kinetic data from cells in different physiological status. A number of specific glycosidases are presently being prepared in this laboratory, including a new enzyme endo-B-N-acetylglucosaminidase-D and a-mannosidase, B-galactosidase, B-N-acetylgluco-saminidase and a-fucosidase. These enzymes will be used to detect structural alterations in the core and protein linkage regions of oligosaccharides of surface glycoproteins. Individual species of glycoprotein will be obtained from growing, density inhibited and transformed cells for comparison.